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. 2016 Jan 24;44(6):2873–2887. doi: 10.1093/nar/gkw040

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© The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 5. — Functional implication of DIS3L2 in miRNA degradation. (A) Efficiency of TUT1 and DIS3L2 knockdowns by western blot. The control siRNA (siCtrl) is targeting the Renilla luciferase. Tubulin serves as loading control. (B) Representative examples of the most abundant miR-27a isoforms detected in one of the libraries treated with the control siRNA and control antimiRNA (antimiR-67) or antimiR-27. Tailed nucleotides are highlighted in purple. The relative abundance of each isoform (%) is given before (antimiR-67) or after (antimiR-27) TDMD induction by the transfection of the corresponding antimiRNAs. The red color indicates a decrease and the green an increase in abundance of the respective sequence. (C) Distribution in percentage of miR-27a tailed and trimmed isoforms as assessed by small RNAs deep-sequencing of cells treated with the indicated siRNA and antimiRNA oligonucleotides. The ‘0’ indicates the wild-type (WT) isoform of 21 nt. The sum of all trimmed (≤-2) or tailed (≥+2) sequences is indicated. (D) The efficiency of the trimming was evaluated in each knockdown condition and antimiRNA treatment. The graph represents the ratio of miR-27a (purple) or miR-27b (blue) trimmed versus WT (‘0’ or mature sequence) reads after TDMD induction (antimiR-27) normalized to those observed without TDMD (antimiR-67) using the values shown in C. The values were arbitrarily set at 1 in cells transfected with the control siRNA (siCtrl). (E) The efficiency of the tailing in each knockdown condition was analyzed as explained in D, except that the graph represents the ratio of tailed isoforms versus WT (‘0’) reads.